124-Color Super-resolution Imaging by Engineering DNA-PAINT Blinking Kinetics
124-Color Super-resolution Imaging by Engineering DNA-PAINT Blinking Kinetics
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DOI：
10.1021/acs.nanolett.9b00508
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发表时间：
2019-04-01
期刊：
影响因子：
10.8
通讯作者：
Jungmann, Ralf
中科院分区：
文献类型：
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作者：
Wade, Orsolya K.;Woehrstein, Johannes B.;Jungmann, Ralf
光学超分辨率技术达到了前所未有的空间分辨率低至几纳米。然而，用于同时检测数百种分子种类的有效多路复用策略仍然是难以捉摸的。在这里，我们介绍了一种全新的方法，多路复用超分辨率显微镜设计的闪烁行为的目标与工程结合频率和持续时间的DNA-PAINT。我们使用DNA折纸结构在计算机和体外分析了这种动力学条形码方法，显示了细胞中多重RNA和蛋白质检测的适用性，并最终在几分钟内通过实验证明了124重超分辨率成像。
Optical super-resolution techniques reach unprecedented spatial resolution down to a few nanometers. However, efficient multiplexing strategies for the simultaneous detection of hundreds of molecular species are still elusive. Here, we introduce an entirely new approach to multiplexed super-resolution microscopy by designing the blinking behavior of targets with engineered binding frequency and duration in DNA-PAINT. We assay this kinetic barcoding approach in silico and in vitro using DNA origami structures, show the applicability for multiplexed RNA and protein detection in cells, and finally experimentally demonstrate 124-plex super-resolution imaging within minutes.